Abstract
Various molecules such as cytokines and anticancer drugs, as well as factor deprivation, rapidly induce apoptosis (programmed cell death)1,2, which is morphologically characterized by cell shrinkage and the blebbing of plasma membranes and by nuclear condensation3,4. Caspases, particularly caspase 3, are proteases that are activated during apoptosis and which cleave substrates such as poly(ADP-ribose) polymerase, actin, fodrin, and lamin5,6. Apoptosis is also accompanied by the internucleosomal degradation of chromosomal DNA7,8,9. In the accompanying Article10, wehave identified and molecularly cloned a caspase-activated deoxyribonuclease (CAD) and its inhibitor (ICAD). Here we show that caspase 3 cleaves ICAD and inactivates its CAD-inhibitory effect. We identified two caspase-3 cleavage sites in ICAD by site-directed mutagenesis. When human Jurkat cells were transformed with ICAD-expressing plasmid, occupation of the receptor Fas, which normally triggers apoptosis, did not result in DNA degradation. The ICAD transformants were also resistant to staurosporine-induced DNA degradation, although staurosporine still killed the cells by activating caspase. Our results indicate that activation of CAD downstream of the caspase cascade is responsible for internucleosomal DNA degradation during apoptosis, and that ICAD works as an inhibitor of this process.
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23 September 2015
Nature 391, 96–99 (1998); doi:10.1038/34214 Recently, it has come to our attention that in Fig. 1a of this Letter, lanes 1 and 5 appear to be duplicated and lanes 6 and 10 appear to be duplicated. It is unclear how this happened. We have repeated the experiment (see Fig. 1 of this Corrigendum) and the results were as described in the original Letter.
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Acknowledgements
We thank R. V. Talanian for the caspase 3 expression plasmid, G. G. Poirier for anti-human poly(ADP-ribose) polymerase, M. A. Blanar for pGEX-2T[128/129], and S. Kumagai for secretarial assistance. This work was supported in parts by Grants-in-Aid from the Ministry of Education, Science, Sports and Culture in Japan.
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Sakahira, H., Enari, M. & Nagata, S. Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391, 96–99 (1998). https://doi.org/10.1038/34214
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DOI: https://doi.org/10.1038/34214
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